Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 May;34(5):1136–1142. doi: 10.1128/jcm.34.5.1136-1142.1996

Genomic analysis of Mycobacterium bovis and other members of the Mycobacterium tuberculosis complex by isoenzyme analysis and pulsed-field gel electrophoresis.

M M Feizabadi 1, I D Robertson 1, D V Cousins 1, D J Hampson 1
PMCID: PMC228969  PMID: 8727890

Abstract

Initially, multilocus enzyme electrophoresis was used to examine genetic relationships among 63 isolates of Mycobacterium bovis and 13 other members of the M. tuberculosis complex. The isolates were divided into five electrophoretic types, with a mean genetic diversity of 0.1. The strains were genetically homogenous, indicating that members of the complex were closely related. This supported the suggestion that they should be considered as subspecies of a single species. Pulsed-field gel electrophoresis (PFGE) was then used to differentiate these isolates, as well as 59 additional isolates of M. bovis from different parts of the world. PFGE differentiated these strains into 63 patterns (53 patterns for M. bovis). Isolates of M. bovis from Western Australia (n = 46) were more homogenous than isolates from other regions. Eight strains were identified in that state, and one predominantly bovine strain was isolated from two human beings and a feral pig. Although M. bovis isolates from different parts of the world had distinct DNA patterns, some were very similar. PFGE is a highly discriminatory technique for epidemiological studies of bovine tuberculosis. For example, it allowed differentiation between isolates of M. bovis cultured from animals in separate outbreaks of tuberculosis, it suggested the transmission of infection between certain properties, and it demonstrated the existence of multiple infections with different strains at certain farms.

Full Text

The Full Text of this article is available as a PDF (301.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Collins C. H., Grange J. M. The bovine tubercle bacillus. J Appl Bacteriol. 1983 Aug;55(1):13–29. doi: 10.1111/j.1365-2672.1983.tb02643.x. [DOI] [PubMed] [Google Scholar]
  2. Collins D. M., De Lisle G. W. DNA restriction endonuclease analysis of Mycobacterium bovis and other members of the tuberculosis complex. J Clin Microbiol. 1985 Apr;21(4):562–564. doi: 10.1128/jcm.21.4.562-564.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collins D. M., De Lisle G. W. DNA restriction endonuclease analysis of Mycobacterium tuberculosis and Mycobacterium bovis BCG. J Gen Microbiol. 1984 Apr;130(4):1019–1021. doi: 10.1099/00221287-130-4-1019. [DOI] [PubMed] [Google Scholar]
  4. Collins D. M., Erasmuson S. K., Stephens D. M., Yates G. F., De Lisle G. W. DNA fingerprinting of Mycobacterium bovis strains by restriction fragment analysis and hybridization with insertion elements IS1081 and IS6110. J Clin Microbiol. 1993 May;31(5):1143–1147. doi: 10.1128/jcm.31.5.1143-1147.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Collins D. M., de Lisle G. W., Collins J. D., Costello E. DNA restriction fragment typing of Mycobacterium bovis isolates from cattle and badgers in Ireland. Vet Rec. 1994 Jun 25;134(26):681–682. doi: 10.1136/vr.134.26.681. [DOI] [PubMed] [Google Scholar]
  6. Cousins D. V., Francis B. R., Gow B. L., Collins D. M., McGlashan C. H., Gregory A., Mackenzie R. M. Tuberculosis in captive seals: bacteriological studies on an isolate belonging to the Mycobacterium tuberculosis complex. Res Vet Sci. 1990 Mar;48(2):196–200. [PubMed] [Google Scholar]
  7. Cousins D. V., Williams S. N., Ross B. C., Ellis T. M. Use of a repetitive element isolated from Mycobacterium tuberculosis in hybridization studies with Mycobacterium bovis: a new tool for epidemiological studies of bovine tuberculosis. Vet Microbiol. 1993 Oct;37(1-2):1–17. doi: 10.1016/0378-1135(93)90178-a. [DOI] [PubMed] [Google Scholar]
  8. Dolin P. J., Raviglione M. C., Kochi A. Global tuberculosis incidence and mortality during 1990-2000. Bull World Health Organ. 1994;72(2):213–220. [PMC free article] [PubMed] [Google Scholar]
  9. Essey M. A., Koller M. A. Status of bovine tuberculosis in North America. Vet Microbiol. 1994 May;40(1-2):15–22. doi: 10.1016/0378-1135(94)90043-4. [DOI] [PubMed] [Google Scholar]
  10. Kristjansson M., Green P., Manning H. L., Slutsky A. M., Brecher S. M., von Reyn C. F., Arbeit R. D., Maslow J. N. Molecular confirmation of bacillus Calmette-Guérin as the cause of pulmonary infection following urinary tract instillation. Clin Infect Dis. 1993 Aug;17(2):228–230. doi: 10.1093/clinids/17.2.228. [DOI] [PubMed] [Google Scholar]
  11. Lygren S. T., Closs O., Bercouvier H., Wayne L. G. Catalases, peroxidases, and superoxide dismutases in Mycobacterium leprae and other mycobacteria studied by crossed immunoelectrophoresis and polyacrylamide gel electrophoresis. Infect Immun. 1986 Dec;54(3):666–672. doi: 10.1128/iai.54.3.666-672.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lévy-Frébault V. V., Portaels F. Proposed minimal standards for the genus Mycobacterium and for description of new slowly growing Mycobacterium species. Int J Syst Bacteriol. 1992 Apr;42(2):315–323. doi: 10.1099/00207713-42-2-315. [DOI] [PubMed] [Google Scholar]
  13. Lévy-Frébault V. V., Thorel M. F., Varnerot A., Gicquel B. DNA polymorphism in Mycobacterium paratuberculosis, "wood pigeon mycobacteria," and related mycobacteria analyzed by field inversion gel electrophoresis. J Clin Microbiol. 1989 Dec;27(12):2823–2826. doi: 10.1128/jcm.27.12.2823-2826.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Musser J. M., Kroll J. S., Moxon E. R., Selander R. K. Clonal population structure of encapsulated Haemophilus influenzae. Infect Immun. 1988 Aug;56(8):1837–1845. doi: 10.1128/iai.56.8.1837-1845.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nei M. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics. 1978 Jul;89(3):583–590. doi: 10.1093/genetics/89.3.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Olson E. S., Forbes K. J., Watt B., Pennington T. H. Population genetics of Mycobacterium tuberculosis complex in Scotland analysed by pulsed-field gel electrophoresis. Epidemiol Infect. 1995 Feb;114(1):153–160. doi: 10.1017/s0950268800052006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Selander R. K., Caugant D. A., Ochman H., Musser J. M., Gilmour M. N., Whittam T. S. Methods of multilocus enzyme electrophoresis for bacterial population genetics and systematics. Appl Environ Microbiol. 1986 May;51(5):873–884. doi: 10.1128/aem.51.5.873-884.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith J. M., Smith N. H., O'Rourke M., Spratt B. G. How clonal are bacteria? Proc Natl Acad Sci U S A. 1993 May 15;90(10):4384–4388. doi: 10.1073/pnas.90.10.4384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stanford J. L., Grange J. M. The meaning and structure of species as applied to mycobacteria. Tubercle. 1974 Jun;55(2):143–152. doi: 10.1016/0041-3879(74)90008-7. [DOI] [PubMed] [Google Scholar]
  20. Thierry D., Cave M. D., Eisenach K. D., Crawford J. T., Bates J. H., Gicquel B., Guesdon J. L. IS6110, an IS-like element of Mycobacterium tuberculosis complex. Nucleic Acids Res. 1990 Jan 11;18(1):188–188. doi: 10.1093/nar/18.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zhang Y., Mazurek G. H., Cave M. D., Eisenach K. D., Pang Y., Murphy D. T., Wallace R. J., Jr DNA polymorphisms in strains of Mycobacterium tuberculosis analyzed by pulsed-field gel electrophoresis: a tool for epidemiology. J Clin Microbiol. 1992 Jun;30(6):1551–1556. doi: 10.1128/jcm.30.6.1551-1556.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. van Soolingen D., Hermans P. W., de Haas P. E., Soll D. R., van Embden J. D. Occurrence and stability of insertion sequences in Mycobacterium tuberculosis complex strains: evaluation of an insertion sequence-dependent DNA polymorphism as a tool in the epidemiology of tuberculosis. J Clin Microbiol. 1991 Nov;29(11):2578–2586. doi: 10.1128/jcm.29.11.2578-2586.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. van Soolingen D., Qian L., de Haas P. E., Douglas J. T., Traore H., Portaels F., Qing H. Z., Enkhsaikan D., Nymadawa P., van Embden J. D. Predominance of a single genotype of Mycobacterium tuberculosis in countries of east Asia. J Clin Microbiol. 1995 Dec;33(12):3234–3238. doi: 10.1128/jcm.33.12.3234-3238.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. van Soolingen D., de Haas P. E., Haagsma J., Eger T., Hermans P. W., Ritacco V., Alito A., van Embden J. D. Use of various genetic markers in differentiation of Mycobacterium bovis strains from animals and humans and for studying epidemiology of bovine tuberculosis. J Clin Microbiol. 1994 Oct;32(10):2425–2433. doi: 10.1128/jcm.32.10.2425-2433.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES